Certain natural products, such as various enzymes, herbs, and plant extractives are used in medicine, food technology, and industry to perform critical functions. For example a range of "crude enzyme" products are used as the basis for preparing oral antiinflammatory drugs. Bromelain, papain, amylase, pancreatin, chymotrypsin, a protease from Serratia marcescens and egg white lysozyme are all occasionally effective as oral antiinflammatory agents. Unfortunately even though all of these enzymes may be rigidly standardized for the named enzyme, the pharmacological effectiveness of different batches of these enzymes varies greatly. Obviously all of the members of this wide range of enzymes contains some pharmacologically active ingredient other than the named enzyme. If the nature and composition of this unknown ingredient could be identified and standardized, then preparations could be made which would be more effective than the improperly standardized present products and new applications could be developed, since the new product would be reliable. This is one of the objectives of the present invention.
In food technology three of the most important applications of "enzymes" are the "chillproofing" of beer, the in vivo and in vitro tenderization of meat, and the preparation of "instant" cooking cereals. Although proteases are used in all of these applications, in no case is there any correlation between proteolytical activity and effectiveness of the preparation. Swift & Co. after much research on their "proten" process found that no known enzyme test could distinguish between effective and non-effective batches of enzymes. On the whole they found that the most proteolytically active samples of papain were the least effective meat tenderizing agents. Since each assay on living animals cost them (1963 figures) $25,000, the importance of this problem can readily be appreciated. The Cream of Wheat Co. in a patented process attempted to use proteases to prepare "instant cooking" cereal. They had so much difficulty in getting reproducible results, even though they purchased enzymes having exactly the same specifications from the same company, that for many years they discontinued using the process. Formulators of beer "chill-proofing" preparations have long realized that each new batch of enzyme which they bought was a gamble. They have never been able to formulate their products on the basis of "proteolytic activity." Instead the better formulators all rely solely upon laboratory use-tests in which the batches of enzymes are evaluated on the basis of their "chill-proofing" ability. If a simple, meaningful assay could be devised which would give a proper evaluation of performance, their costs would be greatly reduced. There have also been some interesting potential application of "enzymes" which have never developed commercially since one batch of "enzyme" might perform satisfactorily whereas the next batch from the same company and having precisely the same specifications would be worthless. The production of better flavored cocoa and vanilla are two examples of such applications.
Three things convinced me that the primary active ingredient in all these "enzymes" was something other than an enzyme; (1) the enzyme rationale for pharmacological activity was completely incompatible with accepted physiological and anatomical data, (2) both clinical and laboratory data showed a lack of correlation between enzymatic activity, and pharmacological action, and (3) certain (but not all) boiled enzyme preparations were as active as were the unheated enzyme preparations. Although certain clues quickly suggested to me that the active ingredient was a small molecule rather than a protein or large polypeptide, working out the details by which this small molecule is produced proved to be a difficult problem. The final solution of this problem now makes it possible to both describe useful methods for producing products containing the active ingredient and to suggest many new combinations and applications for the future.